Mediators of inflammation: unravelling the role of methionine and tryptophan during infection

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Microalgal biomasses have potential as ingredients in microdiets for Senegalese sole (Solea senegalensis) post-larvae

Publication . Peixoto, Diogo; Pinto, Wilson; Gonçalves, Ana Teresa; Machado, Marina; Reis, Bruno; Silva, Joana; Navalho, Joao; Dias, Jorge; Conceicao, Luis; Costas, Benjamin

Senegalese sole (Solea senegalensis) production presents several nutritional challenges, making this species a good candidate to study the dietary potential of bioactive compounds. Since proper nutrition plays a fundamental role in fish biology, it is necessary to further investigate species-specific and well-balanced diets in order to improve Senegalese sole juvenile farming. Algae have antioxidant properties, high-quality dietary protein, and are a source of bioactive compounds. This study evaluates the effects of dietary microalgal inclusion in both health status and growth performance of Senegalese sole post-larvae. Individuals 41 days after hatching (DAH) were randomly distributed among 12 tanks and four experimental diets were randomly distributed by triplicate groups. A basal diet served as CTRL and the experimental diets were formulated to include 3% of each of the algal biomass (CHLO, Chlorella sp. from heterotrophic production; PHAEO, Phaeodactylum sp.; and NANNO, Nannochloropsis sp.). At 50 DAH, 20 post-larvae/tank were collected and homogenized for analysis of immune and oxidative status, and at 61 DAH the total length, dry weight, and survival were assessed. No changes were observed in survival and total length of individuals, post-larvae fed NANNO, and CHLO dietary treatments increased dry weight at 61 DAH compared with those fed CTRL. While post-larvae immune status was apparently not altered by dietary treatments at 50 DAH, the total glutathione content decreased in fish fed PHAEO and CHLO dietary treatments compared to control diet. The observed results on improvement of growth performance without adverse effects on the immune status and decrease of endogenous total glutathione point to the fact that Nannochloropsis sp., Phaeodactylum sp., and Chlorella sp. could work as potential candidates for inclusion in microdiets for Senegalese sole.

2021Journal article

Restricted access

Dietary methionine supplementation improves the European seabass (Dicentrarchus labrax) immune status following long-term feeding on fishmeal-free diets

Publication . Machado, Marina; Engrola, Sofia; Colen, R.; Conceicao, Luis E. C.; Dias, Jorge; Costas, Benjamin

Methionine is a limiting amino acid (AA) in fish diets, particularly in those containing high levels of plant protein (PP), and is key in the immune system. Accordingly, outcome on the fish immune mechanisms of methionine-deficient and methionine-supplemented diets within the context of 0 % fishmeal formulation, after a short and prolonged feeding period, was studied in European seabass (Dicentrarchus labrax). For this, seabass juveniles were fed a (i) fishmeal-free diet, meeting AA requirements, but deficient in methionine (MET0·65); (ii) as control, the MET0·65 supplemented with L-methionine at 0·22 % of feed weight (CTRL); (iii) two diets, identical to MET0·65 but supplemented at 0·63 and 0·88 % of feed weight of L-methionine (MET1·25 and MET1·5, respectively); and (iv) a fishmeal-based diet (FM), as positive control. After 2 and 12 weeks of feeding, blood and plasma were sampled for leucocyte counting and humoral parameter assays and head-kidney collected for gene expression. After 2 weeks of feeding, a fishmeal-free diet supplemented with methionine led to changes in the expression of methionine- and leucocyte-related genes. A methionine immune-enhancer role was more evident after 12 weeks with an increased neutrophil percentage and a decreased expression of apoptotic genes, possibly indicating an enhancement of fish immunity by methionine dietary supplementation. Furthermore, even though CTRL and FM present similar methionine content, CTRL presented a reduced expression of several immune-related genes indicating that in a practical PP-based diet scenario, the requirement level of methionine for an optimal immune status could be higher.

2020-11Journal article

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Two probiotic candidates of the genus psychrobacter modulate the immune response and disease resistance after experimental infection in turbot (Scophthalmus maximus, Linnaeus 1758)

Publication . Wuertz, Sven; Beça, Filipa; Kreuz, Eva; Wanka, Konrad M.; Azeredo, Rita; Machado, Marina; Costas, Benjamin

Probiotic bacteria are a recognized alternative to classical methods of disease prophylaxis and therapy. We tested the effects of their application on the immune reaction in juvenile turbot. To prevent digestion of the probiotics, rectal administration was applied to maximise colonization, by-passing digestion in the stomach. The application of Psychrobacter nivimaris and Psychrobacter faecalis showed beneficial effects on the inflammatory response and disease resistance after infection with the common pathogen Tenacibaculum maritimum. Treatment with P. nivimaris and P. faecalis resulted in 0% and 8% mortalities post-infection, while in the treatment control, an elevated mortality of 20% was observed. In the challenge controls (no infection), no mortalities were observed during the entire experimental period. After an experimental infection, mRNA expression of selected immune markers (mhc II alpha, il-1 beta, tcr, tgf beta and tnf alpha) were determined by RT-QPCR at 0, 1 and 5 days post-infection (dpi). At 0 dpi, gene expression was comparable between the treatments and the treatment control, suggesting that probiotics did not act via immune stimulation of the host. At 1 dpi, all genes were up-regulated in the treatment control but not in the probiotic groups, indicating that the infection in probiotic-treated fish developed at a less severe level. At 5 dpi, mRNA expression returned to baseline levels. As a conclusion, the native probiotic candidates P. nivimaris and P. faecalis improved survival, whereas, in the control, mortality increased and expression of the immune markers was up-regulated post infection. This highlights a potential application of P. nivimaris and P. faecalis in disease prophylaxis, but further research is needed.

2023-03Journal article

Open access